Challenges to Explore Genus Streptomyces in Ethiopia-A Mini Review

Genus Streptomyces is gram-positive bacteria that grow in various environments. It has plentiful biotechnological attributes on the area of agricultural, bioremediation, biofuel, clinical, food, industrial, medical, pharmaceutical, and veterinary. The aim of the review is to frontward challenges to explore potent Streptomyces species in the case of Ethiopia. There is lack of the extent and quality of the genetic research regarding to genome sequence, bioactive compound discovery, and genetic manipulation. Their functional and structural diversity is not full studied. To find a new Streptomyces species: Culture media formulation and optimization as wells culture-independent method like Next Generation Sequencing approach should incorporate at national level.

Kultur Embrio Tiga Spesies Kopi pada Umur Buah dan Formulasi Media yang Berbeda

<em>Studying the fruit age and proper media formulation is one of the important stages in embryo culture of coffee. The data is highly benefical, especially in saving embryos generated from intra- and inter-species crosses that fall prematurely or experience problems in germination. The aim of this study was to determine the suitable age and media formulation for embryo culture of Arabica, Robusta, and Liberica coffee. The study was conducted at the Tissue Culture Laboratory, Indonesian Industrial and Beverage Crops Research Institute from January 2019 to November 2020. Murashige and Skoog (MS) media with growth regulators adapted to embryonic development were used in this study. The three types of coffee divided into 5 groups, namely pinhead, immature, early mature, almost mature, mature, and used as planting material. The research was designed in a completely randomized design with 10 replications, and media formulation as a treatment. The results showed that embryo culture of the three coffee species was conducted successfully, except for pinhead fruit. The older the cultured fruit, the higher the percentage of germination. There is a difference in germination time between the three coffee species. The medium for embryo culture should be adjusted with the age of the fruit being cultured. Aside from growing embryos, the cultured mature fruit embryos on MS medium given 0.5 mg/l BA can also be used for propagation by utilizing the secondary somatic embryos formation.</em>

Can Culture Media Impact Preimplantation Embryo Aneuploidy?

With continued improvements in blastocyst culture, cell sampling approaches, and genetic analysis platforms, the resulting improvements in embryo development and the resolution and accuracy of chromosome analysis have provided valuable insights into the preimplantation embryo. This includes the impact of in vitro culture conditions on chromosomal dynamics. Specifically, through analysis of embryo aneuploidy and mosaicism, a growing number of reports indicate that rates of chromosomal abnormalities can vary between IVF centers. Because differences in mosaicism reflect mitotic errors, this endpoint analysis suggests that IVF laboratory-controlled variables during embryo development may be influencing chromosome separation and segregation. A growing body of literature suggests that culture media may be one variable influencing preimplantation embryo aneuploidy and mosaicism. However, these data are far from definitive in demonstrating cause-and-effect. Whether reported differences may be due to media formulation, use of sequential media or single-step media, or uninterrupted culture approaches is unknown. Importantly, variables directly impacting media performance and embryo development, including pH, temperature, osmolality, and oxygen concentration, must also be considered and make it difficult to isolate the impact of culture media as the sole factor responsible. These IVF laboratory variables will be reviewed and literature suggesting a possible link to mitotic aneuploidy/mosaicism will be discussed.

In vitro propagation of six selected sugarcane mutant clones through leaf explants

Six mutant clones of sugarcane with high productivity have been produced through tissue culture techniques combined with mutations using gamma-ray irradiation and Ethyl Methane Sulfonate. The six mutant clones have been tested for stability in the field. They are proven to have high productivity and yields, so that they are very potential to be developed as superior varieties. To support the planting material sufficiency of these clones, an efficient propagation method was needed. Media formulations with different physical properties and composition of growth regulators were tested to obtain high seedling propagation rates. The media formulation for callus induction was Murashige dan Skoog (MS) + 3 mg/l 2,4-D + 3 g/l casein hydrolysate + 3% sucrose and for shoot regeneration was MS + 0,5 mg/l BA + 0,1 mg/l IBA + 100 mg/l PVP and 2% sucrose. Shoot proliferation was carried out on MS liquid (1, ½) + (0.3; 0.5 mg/l) BA + 0.1 mg/l IBA + 1 mg/l Kinetin + (0; 0.5 mg/l) GA3+ sucrose 2%. The results showed that callus induction, callus regeneration, and shoot proliferation of sugarcane mutant clones were influenced by the genotype and medium composition. The fastest callus induction was obtained from the MSP-4 clone (5.82 days), and the longest was MSB-7 (8.82 days). The largest callus diameter was obtained from MSB-6 clone on MS medium containing 1 mg/l BA, 100 mg/l PVP, and 2% sucrose. The highest number of shoots was obtained from the MSB-6 clone, while the least number of shoots conducted from the MSB-8 clone. The MSB-8 clones were more difficult to regenerate compared to the others. The best media formulation for shoot proliferation was ½ MS containing 0.5 mg/l BA, 1 mg/l Kinetin, and 0.1 mg/l IBA, while the best formulation for rooting was ½ MS.

Comparable Expressions of Bacillus species on Different Growth Media

An investigation was carried out using Pikovskaya Broth (PKB), Luria Bertani Broth (LBB), and Peptone Water (PW) to analyse growth expressions of constructed Bacillus subtilis sub sp and compared to a commercial Bacillus subtilis RIK 1285. The aim was to determine the effect of carbon, nitrogen and other elements at different variations on the metabolic activities under different conditions. The results obtained showed growth density of 4.1 g/ml at 70oC and 3.1 g/ml at pH 6.0; 3.3 g/ml at 70oC and 2.8 g/ml at pH 6.0; 3.8 g/ml at 60oC and 2.6 g/ml at pH 7.0 from PKB, LBB and PW respectively. The growth density of the commercial strain recorded 3.8 g/ml at 50oC and 2.8 g/ml at pH 7.0; 3.1 g/ml at 50oC and 2.3 g/ml; 3.0 at 50oC and 2.3 at pH respectively. The investigation showed importance and relevance of gene metabolic upgrade on the utilization of multiple nutrients present from one media to another. Keywords: media formulation, microbial reaction, growth promoters, growth density

Vero Cells Gain Renal Tubule Markers in Low-calcium and Magnesium Chemically Defined Media

In this study, a chemically defined, animal component-free media was developed to promote Vero growth in suspension. Key media compounds were screened using Plackett-Burman styled experiments to create a media formulation to support suspension growth. Vero cells remained viable in suspension, but their growth rate was extremely low, conversely, other cell types such as CHO-K1, MDCK and HEK293T were able to grow in single cell suspension in the same media. To investigate the slow growth of Vero cells, RNA-seq analysis was conducted. Vero cells were cultured in three different conditions: adherently in serum-containing medium, adherently in in-house medium, and in suspension in low calcium and magnesium in-house medium. This study illustrates that adherent cells maintain similar gene expression, while the suspension phenotype tends to overexpress genes related to renal tubules.

Optimization of phosphate-limited autoinduction broth for two-stage heterologous protein expression in Escherichia coli

Autoinducible, two-stage protein expression leveraging phosphate-inducible promoters has been recently shown to enable not only high protein titers but also consistent performance across scales from screening systems (microtiter plates) to instrumented bioreactors. However, to date, small-scale production using microtiter plates and shake flasks relies on a complex autoinduction broth (AB) that requires making numerous media components, not all amenable to autoclaving. In this report, the authors develop a simpler media formulation (AB-2) with just a few autoclavable components. AB-2 is robust to small changes in its composition and performs equally, if not better, than AB across different scales. AB-2 will facilitate the adoption of phosphate-limited two-stage protein expression protocols.

Impact of artificial sputum media formulation on Pseudomonas aeruginosa secondary metabolite production

In vitro culture media are being developed to understand how host site-specific nutrient profiles influence microbial pathogenicity and ecology. To mimic the cystic fibrosis (CF) lung environment, a variety of artificial sputum media (ASM) have been created. However, the composition of these ASM vary in the concentration of key nutrients, including amino acids, lipids, DNA, and mucin. In this work, we used feature-based molecular networking (FBMN) to perform comparative metabolomics of Pseudomonas aeruginosa , the predominant opportunistic pathogen infecting the lungs of people with CF, cultured in nine different ASM. We found that the concentration of aromatic amino acids and iron from mucin added to the media contribute to differences in the production of P. aeruginosa virulence-associated secondary metabolites. IMPORTANCE Different media formulations aiming to replicate in vivo infection environments contain different nutrients, which affects interpretation of experimental results. Inclusion of undefined components, such as commercial porcine gastric mucin (PGM), in an otherwise chemically defined medium can alter the nutrient content of the medium in unexpected ways and influence experimental outcomes.